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For genes with alternative splicing isoforms, we retained the longest mRNA for analysis.
In the case of alternative splicing variants, we used the longest mRNA for analysis (although similar results were obtained by analyzing the shortest mRNA, data not shown).
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To address this, we cloned the 5′ UTR from the longest mRNAs for the mouse insulin receptor (mINR) and the mouse insulin-like growth factor receptor-I (IGFR) and created firefly luciferase reporters under the control of these UTRs.
For the genes with alternatively spliced isoforms, the longest mRNA was used for analysis.
The longest mRNA transcript variant for every tumor suppressor gene of interest was scrutinized using RNA22 to identify miR-10b-binding sites.
The remaining 21 polymorphisms are located at the region coding for the longest mRNA (uc001aoq.2). Figure 1A shows the genomic organisation in this region.
The PAS located closest to the 3'end is conserved in all species and represents the signal for the longest mRNA version.
We find that protein accumulation with time per cell is quadratic for proteins with long mRNA life times and approximately linear for a protein with short mRNA lifetime.
AE analysis must be performed specifically for each splice variant, as demonstrated for the short and long mRNA isoforms of dopamine receptor D2 [ 20].
For example, most housekeeping genes have long mRNA half-lives.
Hybrids between heterologous mRNAs, generated by mRNA trans-splicing or transcription of long mRNA across neighbouring loci may provide the cell with the potential for evolution and adaptation by acquiring new functions.
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